Researchers using radar, thermal imaging and a miniature probe believe they have found a lost masterpiece by Leonardo da Vinci hidden behind another fresco in Florence’s city hall for the past 450 years.

Maurizio Seracini, a University of California scientist who was featured in Dan Brown’s bestseller The Da Vinci Code and has been looking for Leonardo’s monumental “Battle of Anghiari” since 1975, announced his findings in Florence on Monday.

“The evidence does suggest that we are searching in the right place,” said Mr Seracini whose quest was driven by his earlier discovery of the words Cerca, trova (He who seeks, finds) painted on a flag in a later 16th century fresco by Giorgio Vasari that is believed to cover the original Leonardo.

Fuelling protests by leading art historians in Italy and elsewhere, Mr Seracini’s team bored six tiny holes through Vasari’s “Battle of Marciano” and inserted a 4mm endoscopic probe through a cavity to take samples of paint behind.

The team reported that the chemical composition of the black material was similar to pigment found in brown glazes on Leonardo’s “Mona Lisa” and “St John the Baptist”, with Mr Seracini noting that Leonardo painted the “Mona Lisa” in Florence between 1503 and 1506, at the same time as the “Battle of Anghiari”. Only 18 of Leonardo’s paintings are known to exist.

Vasari had been ordered by Cosimo the Great, duke of Florence, to refurbish the Hall of the Five Hundred in Florence’s Palazzo Vecchio, now the city hall.

Matteo Renzi – the mayor of Florence who backed the project and has been accused by art historians of seeking publicity at the expense of damaging Vasari’s fresco – told the Financial Times that the team believed Vasari, a great admirer of Leonardo, had disobeyed instructions to destroy the fresco to replace it with a battle scene commemorating Cosimo’s victory over the Republic of Siena.

Instead Vasari may have built a brick wall over Leonardo’s work and painted his fresco on top in 1563. Mr Seracini’s team noted that no other cavities existed behind the five other Vassari frescoes in the hall. The team also found flakes of organic red material which could be lacquer, and a beige material believed to have been applied with a brush.

Leonardo is believed to have completed only the centrepiece of his giant fresco, having experimented with materials and run into trouble with running paints and wax. The most famous copy of that scene, by Peter Paul Rubens, showing horsemen battling over trampled infantry, hangs in the Louvre.

Mr Seracini said they were still in the preliminary stages of research, which is being part sponsored by National Geographic, and a lot more work was needed to solve the mystery.

Tomaso Montanari, history professor at the University of Naples and a fierce critic of the project, has dismissed it as “more like the search for the Holy Grail than real history”. He said anything painted in the Renaissance era could be on the hidden wall, and wanted a “neutral team” of scientists to evaluate the findings.

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